ORIGINAL RESEARCH Evaluation of Pathological Microbes in Root Canals with Pulp Necrosis Atul Anand Bajoria1, Ahmed Ali Alfawzan2, Vardharajula Venkata Ramaiah3, Mohammed Ali Habibullah4, Sabahat Ullah Tareen5, Prashant Babaji6 ABSTRACT Aim: The present study was conducted to evaluate the type of bacteria present in necrotic root canals of permanent teeth. Materials and methods: All 60 participants with infected root canals were made to use 10 mL of mouthwash containing 0.12% chlorhexidine. Access to pulp chamber was established, and the sterile absorbent paper cones were inserted into the canal for 20 seconds. The contaminated paper cones were inoculated in a brain–heart infusion (BHI) agar culture medium and incubated in an oven for 48 hours at 37°C. Results were analyzed statistically with SPSS version 20.0 using Chi-square test and analysis of variance (ANOVA). Results: In root canals with periapical lesions, gram-positive bacilli was present in 50 cases, gram-negative in 48 and yeast cells in 28; while in root canals without periapical lesions, gram-positive bacilli was present in 8. In 16 root canals of chronic apical periodontitis cases, gram-positive bacteria was present in 100%, gram-negative bacteria in 100%, and yeast cells in 20% cases. In 12 cases of periapical granuloma, gram-positive bacteria was present in 98%, gram-negative bacteria in 100%, and yeast cells in 40% cases. In 10 cases of chronic abscess with fistula, gram- positive bacteria was present in 86.2. In six cases of phoenix abscess, gram-positive bacteria was present in 100% and gram-negative bacteria in 100% cases. Conclusion: There is predominantly gram-positive and -negative bacteria as well as the presence of yeast cells in periapical lesions. Clinical significance: Knowledge about the bacterial species in root canal infection helps in its management and in selecting antibiotic. Keywords: Bacteria, Gram positive, Periapical. World Journal of Dentistry (2020): 10.5005/jp-journals-10015-1723 INTRODUCTION 1Department of Oral Medicine, Kalinga Institute of Dental Sciences, The treatment of infected tooth with endodontic treatment ensures KIIT University, Bhubaneswar, Odisha, India removal of pathogenic bacteria. However, sometimes failure of 2Department of Preventive Dentistry, Al-Rass Dental College, Qassim complete elimination from root canal leads to endodontic failure.1 University, Buraida, Kingdom of Saudi Arabia Chemically, the microorganisms can be eliminated using disinfectant. 3Department of Dental Hygiene, College of Applied Health Sciences in Various disinfectants have been introduced in the treatment of Al-Rass, Qassim University, Buraida, Kingdom of Saudi Arabia 4 infected root canals such as NaOCl, H2O2, etc. The choice of irrigants Department of Pedodontics, College of Dentistry, Qassim University, depends upon cause and amount of infection. Thus, an endodontist Buraida, Kingdom of Saudi Arabia should have a thorough knowledge of endodontic microbiota and 5Department of Restorative Dentistry, College of Dentistry, Qassim 2 infectious process and possible remedies to treat it. University, Buraida, Kingdom of Saudi Arabia Bacteria exhibit their effects by releasing toxins and 6Department of Pedodontics, Sharavathi Dental College, Shimoga, lipopolysaccharides which are constituent of the cell wall. The Karnataka, India virulence is bacteria are of low intensity. The occurrence of Corresponding Author: Prashant Babaji, Department of Pedodontics, bacteria in root canals causes pulpal as well as periapical infection. Sharavathi Dental College, Shimoga, Karnataka, India, Phone: +91 Endotoxins may trigger pain in infected root canal conditions 8971096312, e-mail: [email protected] by releasing vasoactive substances and neurotransmitters at How to cite this article: Bajoria AA, Alfawzan AA, Ramaiah VV, et al. nerve endings of periapical tissues. Endotoxins exerts effects by Evaluation of Pathological Microbes in Root Canals with Pulp Necrosis. interacting with monocytes, macrophages, and other cells of the World J Dent 2020;11(3):211–214. immune systems and ultimately with fibroblasts and resulting in the Source of support: Nil release of proinflammatory cytokines, e.g., tumor necrosis factor-α Conflict of interest: None and interleukin-1β.3 Studies have shown the presence of gram-positive, gram- negative, mixed aerobic and anaerobic bacteria that cause MATERiaLS AND METHODS infection. Most common gram-negative anaerobic bacteria species The present study was conducted in the Department of Restorative involves Porphyromonas, Prevotella, Fusobacterium, Tanerella, and Dentistry, College of Dentistry, Qassim University, Kingdom of Saudi Treponema.4,5 Several techniques such as polymerase chain reaction Arabia. It comprised 40 patients of both genders with 60 infected may be useful in detection of microbes. This study was conducted root canals. The ethical approval was taken from institutional ethical to evaluate the type of bacteria present in the necrotic pulp canals committee of College of Dentistry, Qassim University. Informed of permanent teeth. consent was obtained from participants. Molar teeth with necrotic © The Author(s). 2020 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons. org/licenses/by-nc/4.0/), which permits unrestricted use, distribution, and non-commercial reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Pathological Microbes in Root Canals with Pulp Necrosis root canals and asymptomatic teeth were included. All the canals Table 2 and Figure 1 show that in 16 pulp canals of persistent for particular tooth were evaluated. Exclusion criteria were patients apical periodontitis cases, gram-positive microorganism was using antibiotics and who are under medical conditions. The study present in 100%, gram-negative bacteria in 100%, and yeast cells was done by a single-trained investigator. in 20% cases. In 12 cases of periapical granuloma, gram-positive Clinical examination and pulp vitality were performed. Intraoral bacteria was present in 98%, gram-negative bacteria in 100%, radiographs were taken. All subjects were made to use 10 mL of and yeast cells in 40% cases. In 10 cases of persistent abscess with mouthwash containing 0.12% chlorhexidine. After caries removal, fistula, gram-positive microorganism was present in 86.2%, gram- the access to the pulp chamber was established. negative bacteria in 99%, and yeast cells in 80% cases. In eight cases Three to four sterile permeable paper points were inserted of periapical cysts, gram-positive bacteria was present in 100%, into each canal for 20 seconds. These polluted paper pointes were gram-negative bacteria in 100%, and yeast cells in 42% cases. In inserted in a BHI agar culture carrier and incubated in an oven for six cases of phoenix abscess, gram-positive bacteria were present 48 hours at 37°C. in 100% and gram-negative bacteria in 100% cases. The difference The Gram methodology was used. The prepared thin scrub was was significant (p < 0.05). kept for air desiccation. Following this, it was passed by a burner flame for 3–4 times. Then for 4 minutes it was enclosed with ethanol. DISCUSSION Later its surface was covered for 1 minute with crystal violet solution, cleaned with distilled water, and enclosed for 1 minute with Gram The study of periapical and pulpal diseases shows the existence iodide solution. Then it was cleaned again using distilled water. of complex mixed infections. Various studies have been done so Following this, acetone was added on the material till the purple far which depicts the occurrence of gram-positive, gram-negative color fade, in 10 seconds. The cleansed area was enclosed for 1 bacteria, and yeast cells. Microscopic and macroscopic studies 6,7 minute with safranin and cleaned with deionized water. The material have shown the pathological function in pathogenesis. The was positioned in a vertical location on a holder and kept for air-dry. polymicrobial behavior of endodontic infection can be confirmed and classified based on Gram color. The existence of aerobic The tainted lamina was evaluated under 100× magnification using a drop of immersion oil. Dark blue used for gram-positive microorganisms, strict anaerobes, yeasts, and facultative bacteria 8 bacteria and pink red for gram-negative bacteria. Using objective demands complete elimination for successful endodontic therapy. This study was conducted to evaluate the type of bacteria present lens, 100× optical microscope, morphological classification of microorganisms was achieved. in necrotic pulp canals of permanent teeth. Results were analyzed statistically with SPSS version 20.0. Chi- We included 60 root canals with pulp necrosis in 40 patients. square test and ANOVA were used for representation of the results. Only multirooted mandibular permanent molars were considered The p value was considered significant at lesser than 0.05. for the study. We found that in root canals with periapical lesions, gram-positive bacilli was present in 50 cases, gram-negative RESULTS bacteria in 48, and yeast cells in 28; while in root canals without periapical lesions, gram-positive bacilli was present